miR-23b 通过下调 TIMP3 促进角质形成细胞迁移。

MiR-23b Promotes the Migration of Keratinocytes Through Downregulating TIMP3.

机构信息

Department of Plastic and Burn Surgery, West China Hospital, Sichuan University, Chengdu, Sichuan, China.

出版信息

J Surg Res. 2020 Oct;254:102-109. doi: 10.1016/j.jss.2020.03.043. Epub 2020 May 15.

DOI:10.1016/j.jss.2020.03.043

PMID:32422429

Abstract

BACKGROUND

Wound healing is a complex process aiming at repairing the damaged skin. MiR-23b has been reported to be upregulated during wound healing. In this study, we intended to explore the working mechanism of miR-23b during wound healing.

METHODS

Quantitative real-time polymerase chain reaction was performed to detect the enrichment of miR-23b and tissue inhibitor of metalloproteinase-3 (TIMP3) in HaCaT cells. Scratch wound assay was carried out to measure the migration of HaCaT cells. The target of miR-23b was predicted by microT-CDS software, and the combination was confirmed by dual-luciferase reporter assay and RNA immunoprecipitation assay. The abundance of TIMP3 protein was detected by Western blot assay.

RESULTS

The abundance of miR-23b was positively related to the concentration and time of transforming growth factor β1 treatment in HaCaT cells. MiR-23b promoted the migration of keratinocytes. TIMP3 was a direct target of miR-23b and was negatively regulated by miR-23b. TIMP3 inhibited the migration of keratinocytes. MiR-23b accelerated the migration of keratinocytes by downregulating the abundance of TIMP3.

CONCLUSIONS

MiR-23b promoted the migration of keratinocytes partly through reducing the enrichment of TIMP3. MiR-23b might be a promising target for the treatment of wound healing-associated diseases.

摘要

背景

伤口愈合是一个旨在修复受损皮肤的复杂过程。已经有报道称 miR-23b 在伤口愈合过程中上调。在本研究中，我们旨在探索 miR-23b 在伤口愈合过程中的作用机制。

方法

采用实时定量聚合酶链反应检测 HaCaT 细胞中 miR-23b 和组织金属蛋白酶抑制剂 3（TIMP3）的富集。划痕实验检测 HaCaT 细胞的迁移。microT-CDS 软件预测 miR-23b 的靶标，并通过双荧光素酶报告基因实验和 RNA 免疫沉淀实验验证结合。Western blot 检测 TIMP3 蛋白的丰度。

结果

miR-23b 的丰度与 HaCaT 细胞中转化生长因子 β1 的浓度和作用时间呈正相关。miR-23b 促进角质形成细胞的迁移。TIMP3 是 miR-23b 的直接靶标，并受 miR-23b 负调控。TIMP3 抑制角质形成细胞的迁移。miR-23b 通过下调 TIMP3 的丰度加速角质形成细胞的迁移。